急性肝衰竭大鼠载肝细胞生长因子纳米粒子肝细胞移植后有丝分裂指数和Ki-67表达变化及意义

The change and significance of Ki-67 expression following hepatocyte transplantation using hepatocyte growth factor loaded nanoparticles in rats with acute liver failure

目的探讨急性肝衰竭（AIJF）大鼠载肝细胞生长因子（HGF）的聚乳酸-氧-羧甲基壳聚糖（PLA—O—CMC）纳米粒子肝细胞移植后有丝分裂指数和Ki-67抗原的表达变化及意义。方法D-氨基半乳糖腹腔内注射制作大鼠ALF模型，48h后分别将Ⅰ型胶原（Ⅰ组）、PLA—O—CMC纳米粒子（Ⅱ、Ⅳ组）、载HGF的PLA—O—CMC纳米粒子（Ⅲ组）培养的大鼠肝细胞（均为5×10^7个，5ml）移植到ALF大鼠腹腔内。以每日静脉注射HGF10μg／kg×7d（1V组）、5ml RPMI 1640腹腔内注射（V组）作为对照。观察其14d存活率、血清及肝组织HGF浓度、肝组织病理及有丝分裂指数（MI）、Ki-67变化。结果移植后14dⅠ～Ⅴ组ALF大鼠的存活率分别为50．00％、68．75％、81．25％、75．00％、18．75％。各纳米移植组高于Ⅴ组。Ⅲ组3d时肝组织HGF浓度最高，平均为5882．91μg／L。1I组肝组织结构恢复更快，5d时平均MI10．20％0、7d时Ki-67平均标记指数16．8‰，均高于Ⅴ组。结论应用载HGF的PLA-O-CMC纳米粒子培养的大鼠肝细胞腹腔内移植治疗ALF大鼠能促进肝再生相关抗原表达。

Objective To evaluate the change and significance of Ki-67 antigen expression follow- ing hepatocyte transplantation using hepatocyte growth factor （HGF） loaded polylactic acid-O-carboxymeth-ylchitosan （PLA-O-CMC） nanoparticles in rats with acute liver failure （ALF）. Methods ALF models of rats were established by D-galactosamine intraperitoneal injection. Rat hepatocytes type Ⅰ collagen suspension （group Ⅰ ）, rat hepatocytes co-cultured with PLA-O-CMC nanoparticles （groups Ⅱ , Ⅳ ） and rat hepatocytes co-cultured with HGF loaded PLA-O-CMC nanoparticles （group Ⅲ ） （5 × 10^7 cells each group, 5 ml） were transplanted into the abdominal cavity of SD rats at 48 h after D-Gal injection. Intravenous injection of HGF （10μg/kg for 7 days） （group Ⅳ ） and RPMI 1640 injection （group V ） were used as the negative groups. The 14th-day survival rate of rats, pathological change and mitotic index of liver, Ki-67 antigen labeling index of ALF rat livers were observed. Results The 14th-day survival rate in groups Ⅰ -Ⅴ was 50. 00%, 68.75%, 81.25%, 75.00%, 18.75%, and that in nano-transplanted groups was higher than in group V. At the 3rd day, the concentration of HGF in liver tissue of group Ⅲ, average 5882.91 μg/L was the highest. The hepatic lobules structure in group Ⅲ recovered faster. The average mitotic index in group Ⅲ at the 5th day was 10. 20%0 and the average Ki-67 labeling index at the 7th day was 16. 8‰, which were all higher than in group V. Conclusion Intraperitoneal transplantation of HGF loaded PLA-O-CMC nanoparticle-attached hepatocytes for treatment of ALF can promote the liver regenerationassociated antigen expression.